LOCUS pRF-HU. 6336 bp DNA circular
DEFINITION Complementary copy of pRF-HU c.
COMMENT The vector can be used for ectopic integration of any DNA fragment
genome of filamentous fungi.
The vector has been designed for the directional cloning of a single PCR
amplicon, utilizing the USER friendly cloning technology from NEB.
Add the following 5' overhangs to the primers for amplification of the two
specific homologous recombination flanks:
Gene-U1 (LB forward) 5´- GGGTTTAAU - Forward primer sequence
Gene-U2 (LB reverse) 5'- GGACTTAAU - Reverse primer sequence
Digestion of vector prior to cloning:
1. Digest over night with PacI
2. Digest for 2 hours with Nt.Bbv.C IB (nicking enzyme)
Transformation of the vector into the fungi:
The vector is originate designed for Agrobacterium tumefaciens mediated
transformation, but can also be used for protoplast transformation